The viability of cryopreserved PBPC depends on the DMSO concentration and the concentration of nucleated cells in the graft

Liseth, Knut; Abrahamsen, Jenny Foss; Bjørsvik, Solfrid; Grøttebø, K; Bruserud, Øystein

doi:10.1080/14653240500238251

Cited by 51 publications

(47 citation statements)

References 19 publications

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“…Recently, there have been several reports suggesting that cells can be successfully cryopreserved with minimal concentrations of DMSO [39][40][41][42][43][44][45][46][47][48][49][50][51]. For example, Zhao et al [46] showed that the use of 5% or 10% DMSO in combination with either 20% or 70% serum produced similar results during cryopreservation of fetal human liver CD34 + cells.…”

Section: Discussionmentioning

confidence: 68%

“…Abrahamsen et al [49] showed that cryopreserving peripheral blood progenitor cells (PBPC) with 5% DMSO instead of 10% DMSO produced improved CD34 + cell viability, while Stiff et al [50] developed an alternative cryopreservation protocols for hematopoietic stem cells using 5% DMSO in conjunction with 6% hydroxyethyl starch (HES). Furthermore, Liseth et al [51] concluded that the post-thaw survival for CD34 + was almost identical when frozen with either 4% or 5% DMSO but further lowering the concentration of DMSO to 2% had an adverse effect on post-thaw cell survival.…”

Section: Discussionmentioning

confidence: 99%

“…Although DMSO is regarded as relatively nontoxic, the clinical use of frozen/thawed cells treated with DMSO can cause many adverse effects and toxic reactions [7][8][9][10][11][12][13][14]. It has also been reported that DMSO is not only cytotoxic but it also induces differentiation of stem cells to cardiac or neuronal like cells when added to the cell culture medium [15,16].…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Thirumala

Gimble

Devireddy

2010

Stem Cells and Development

102

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Developing effective techniques for the cryopreservation of human adipose-derived adult stem cells (ASCs) could increase the usefulness of these cells in tissue engineering and regenerative medicine. To this end, we investigated the post-freeze/thaw viability and apoptotic behavior of Passage 1 (P1) adult stem cells (ASCs) in 11 different media: (i) the traditional media containing Dulbecco's modifi ed Eagle's medium (DMEM) with 80% fetal calf serum (FCS) and 10% dimethyl sulfoxide (DMSO), (ii) DMEM with 80% human serum (HS) and 10% DMSO, (iii) DMEM with 1% methyl cellulose (MC) and 10% of either HS or FCS or DMSO, and (iv) DMEM with 0%, 2%, 4%, 6%, 8%, or 10% DMSO. Approximately 1 mL (10 6 cells/mL) of P1 ASCs were frozen overnight in a −80°C freezer and stored in liquid nitrogen for 2 weeks before being rapidly thawed in a 37°C water bath (1-2 min of agitation), resuspended in culture media, and seeded in separate wells of a 6-well plate for a 24-h incubation period at 37°C. After 24 h, the thawed samples were analyzed by bright-fi eld microscopy and fl ow cytometry. The results suggest that the absence of DMSO (and the presence of MC) signifi cantly increases the fraction of apoptotic and/or necrotic ASCs. However, the percentage of viable cells obtained with 2% DMSO and DMEM was comparable with that obtained in freezing media with 10% DMSO and 80% serum (HS or FCS), that is, ~84% ± 5% and ~84% ± 8%, respectively. Adipogenic and osteogenic differentiation behavior of the frozen thawed cells was also assessed using histochemical staining. Our results suggest that post-thaw ASC viability, adipogenic and osteogenic differentiability can be maintained even when they are frozen in the absence of serum but with a minimal concentration of 2% DMSO in DMEM.

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Section: Discussionmentioning

confidence: 68%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Thirumala

Gimble

Devireddy

2010

Stem Cells and Development

102

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“…It has been notably shown that cytotoxicity tolerance of DMSO is evidently much higher in stem cells rather than in normal differentiated ones. It is worth mentioning that differentiated cells can tolerate optimal DMSO concentration up to 0.5%; however, peripheral blood mononuclear cells and MSCs are shown to tolerate DMSO even till 5% [34,35]. When carefully observed, these cells were found to have enormous tolerance for potent cytotoxic agents DMSO.…”

Section: Cell Proliferation and Cytotoxicity Assays Revealed Stemnessmentioning

confidence: 99%

Basal Expression of Pluripotency-Associated Genes Can Contribute to Stemness Property and Differentiation Potential

Dadheech

Srivastava

Belani

et al. 2013

Stem Cells and Development

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Pluripotency and stemness is believed to be associated with high Oct-3/4, Nanog, and Sox-2 (ONS) expression. Similar to embryonic stem cells (ESCs), high ONS expression eventually became the measure of pluripotency in any cell. The threshold expression of ONS genes that underscores pluripotency, stemness, and differentiation potential is still unclear. Therefore, we raised a question as to whether pluripotency and stemness is a function of basal ONS gene expression. To prove this, we carried out a comparative study between basal ONS expressing NIH3T3 cells with pluripotent mouse bone marrow mesenchymal stem cells (mBMSC) and mouse ESC. Our studies on cellular, molecular, and immunological biomarkers between NIH3T3 and mBMSC demonstrated stemness property of undifferentiated NIH3T3 cells that was similar to mBMSC and somewhat close to ESC as well. In vivo teratoma formation with all three germ layer derivatives strengthen the fact that these cells in spite of basal ONS gene expression can differentiate into cells of multiple lineages without any genetic modification. Conclusively, our novel findings suggested that the phenomenon of pluripotency which imparts ability for multilineage cell differentiation is not necessarily a function of high ONS gene expression.

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“…[42][43][44][45][46][47][48][49] It has also been reported that DMSO is not only cytotoxic but it also induces differentiation of stem cells to cardiac or neuronal like cells when added to the cell culture medium. 50,51 Therefore, it is of vital necessity to reduce the toxicity by the removal of DMSO prior to clinical use.…”

Section: Clinical Banking Of Adipose Derived Adult Stem Cells (Ascs)mentioning

confidence: 99%

Clinical grade adult stem cell banking

2009

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The viability of cryopreserved PBPC depends on the DMSO concentration and the concentration of nucleated cells in the graft

Cited by 51 publications

References 19 publications

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

Basal Expression of Pluripotency-Associated Genes Can Contribute to Stemness Property and Differentiation Potential

Clinical grade adult stem cell banking

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